A robust, highly scalable whole-transcriptome analysis (RNA-Seq) solution for a variety of species and sample types, including human, mouse, and formalin-fixed, paraffin-embedded (FFPE) tissue.
Assay time
Hands-on time
Input quantity
The products previously known as TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Human/Mouse/Rat (Cat. No. RS-122-2201, RS-122-2202, and RS-122-2203) and TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Cat. No. RS-122-2301, RS-122-2302, and RS-122-2303) are now called TruSeq Stranded Total RNA Human/Mouse/Rat and TruSeq Stranded Total RNA Gold, respectively. The product configuration has changed. You'll need to purchase the library prep and index adapter products separately (instead of in a kit).
TruSeq Stranded Total RNA provides a clear view of the transcriptome with a streamlined, cost-efficient, and scalable solution for total RNA analysis with sequencing.
Compatible with a wide range of samples, including low-quality DNA/RNA and formalin-fixed, paraffin-embedded (FFPE) samples, TruSeq Stranded Total RNA couples all of the benefits of TruSeq RNA technology with Ribo-Zero ribosomal RNA reduction chemistry. The product enables analysis of coding and multiple forms of noncoding RNA with precise measurement of strand orientation, uniform coverage, and high-confidence discovery of features such as alternative transcripts, gene fusions, and allele-specific expression.
TruSeq Stranded Total RNA is designed for human, mouse and rat sequences, but may be applicable to a variety of eukaryotic species.
TruSeq Stranded Total RNA is available in multiple versions to fit specific study design needs:
Prepare libraries for Illumina sequencing in less than a day. Illumina Qualified Methods are available on a range of automation platforms through our partners.
| Assay time | 11.5 hr |
|---|---|
| Automation capability | Liquid handling robots |
| Automation details | Explore available automation methods |
| Content specifications | Captures coding RNA plus multiple forms of non-coding RNA |
| Description | For whole-transcriptome sequencing studies. The Total RNA version depletes cytoplasmic rRNA while the Total RNA Gold version depletes cytoplasmic and mitochondrial rRNA. |
| Hands-on time | 5.5 hr |
| Input quantity | 0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization. |
| Instruments | NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System |
| Mechanism of action | Bead-based rRNA depletion, cDNA synthesis, and PCR |
| Method | Whole-transcriptome sequencing |
| Multiplexing | Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together, High-throughput kits: Prepare 96 indexed samples using either combinatorial dual indexes or unique dual indexes |
| Nucleic acid type | RNA |
| Specialized sample types | FFPE tissue |
| Species category | Mouse, Human, Rat |
| Species details | rRNA Removal: Depletes human, mouse, and rat rRNA |
| Technology | Sequencing |
| Variant class | Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants |
TruSeq Stranded Total RNA
| Instrument | Recommended number of samples | Read length |
|---|---|---|
| NextSeq 550 System | RNA profiling: 6-20 samples per run (based on 20 million reads per sample) |
2 x 75 bp |
| NovaSeq 6000 System | RNA profiling (samples per run, dual flow cell): S1: 160, S2: 320, S4: 768. Limited by index combinations (dual). Based on 20 million reads. |
≤ 2 × 100 bp |
RNA-Seq uses next-generation sequencing to analyze expression across the transcriptome, enabling scientists to detect known or novel features and quantify RNA.
Low-Quality/FFPE RNA-Seq Solutions
Find FFPE RNA sequencing solutions that yield high-quality results from degraded FFPE tissues and other low-quality samples.
| TruSeq Stranded Total RNA | Illumina Stranded Total RNA Prep with Ribo-Zero Plus or Ribo-Zero Plus Microbiome | |
|---|---|---|
| Assay time | 11.5 hr | ~7 hr |
| Automation capability | Liquid handling robots | Liquid handling robots |
| Automation details | Explore available automation methods | |
| Content specifications | Captures coding RNA plus multiple forms of non-coding RNA | Captures coding RNA plus multiple forms of non-coding RNA |
| Description | For whole-transcriptome sequencing studies. The Total RNA version depletes cytoplasmic rRNA while the Total RNA Gold version depletes cytoplasmic and mitochondrial rRNA. | For the preparation of whole-transcriptome sequencing-ready libraries from low RNA inputs and low-quality samples that supports a wide range of RNA-Seq applications. Includes Ribo-Zero Plus for single-tube depletion of rRNA and globin RNA from multiple species to facilitate rich transcriptome analyses. |
| Hands-on time | 5.5 hr | < 3 hr |
| Input quantity | 0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization. | 1–1000 ng total RNA for standard-quality RNA samples. 10 ng total RNA minimum recommended for optimal performance and FFPE samples |
| Instruments | NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System | NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System |
| Mechanism of action | Bead-based rRNA depletion, cDNA synthesis, and PCR | Enzymatic rRNA depletion, ligation-based addition of adapters and indexes |
| Method | Whole-transcriptome sequencing | Whole-transcriptome sequencing, Metatranscriptome sequencing |
| Multiplexing | Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together, High-throughput kits: Prepare 96 indexed samples using either combinatorial dual indexes or unique dual indexes | Up to 384 Unique Dual Indexes (UDIs) |
| Nucleic acid type | RNA | RNA |
| Specialized sample types | FFPE tissue | Microbiome samples, Low-input samples, FFPE tissue |
| Species category | Mouse, Human, Rat | Mouse, Human, Rat, Bacteria |
| Species details | rRNA Removal: Depletes human, mouse, and rat rRNA | Ribo-Zero Plus depletes abundant transcripts from multiple species including: human cytoplasmic and mitochondrial rRNA, mouse rRNA, rat rRNA, bacteria (E. coli and B. subtilis) rRNA, and human beta globin transcripts Ribo-Zero Plus Microbiome depletes abundant transcripts from multiple species including: those depleted in Ribo-Zero Plus as well as diverse microbial species and ATCC MSA-2002, MSA-2005, and MSA-2006 |
| Technology | Sequencing | Sequencing |
| Variant class | Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants | Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants |
Library Prep and Array Kit Selector
Find the right sequencing library preparation kit or microarray for your needs. Filter by method, species, and more. Compare, share, and order kits.
Consistent, precise measurement of RNA abundance is reflected by high reproducibility between technical replicates. These technical replicates of FFPE tissue show high concordance, indicating robust library prep performance. Axes are log2(FPKM). R2 value is shown.
TruSeq Stranded Total RNA gives excellent coverage across the top 1000 expressed transcripts in both fresh-frozen (FF, top) and FFPE (bottom) tumor and matched normal breast tissue, with > 98% aligned stranded reads. X-axis: position along transcript, Y-axis: percent coverge of combined reads.
TruSeq® Stranded Total RNA Library Prep Human/Mouse/Rat (48 Samples)
20020596
Includes RiboZero beads to deplete rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA , and reagents for preparing 48 samples. Purchase index adapters and Purification Beads separately.
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TruSeq® Stranded Total RNA Library Prep Human/Mouse/Rat (96 Samples)
20020597
Includes RiboZero beads to deplete rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA , and reagents for preparing 48 samples. Purchase index adapters and Purification Beads separately.
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TruSeq Stranded Total RNA Library Prep Gold (48 Samples)
20020598
Includes RiboZero beads to deplete rRNA and mitochondrial rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA, and reagents for preparing 48 samples. Purchase index adapters separately. Purchase index adapters and Purification Beads separately.
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TruSeq® Stranded Total RNA Library Prep Gold (96 Samples)
20020599
Includes RiboZero beads to deplete rRNA and mitochondrial rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA, and reagents for preparing 48 samples. Purchase index adapters separately. Purchase index adapters and Purification Beads separately.
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TruSeq RNA Single Indexes Set A (12 Indexes, 48 Samples)
20020492
Includes 12 of 24 indexes (Indexes: 2, 4, 5, 6, 7, 12, 13, 14, 15, 16, 18, 19) for preparing 48 individual RNA samples.
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TruSeq RNA Single Indexes Set B (12 Indexes, 48 Samples)
20020493
Includes 12 of 24 indexes (Indexes: 1, 3, 8, 9, 10, 11, 20, 21, 22, 23, 25, 27) for preparing 48 individual RNA samples.
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IDT for Illumina – TruSeq RNA UD Indexes v2 (96 Indexes, 96 Samples)
20040871
Includes 96, 8 bp indexes sufficient for labeling 96 samples and is compatible with TruSeq Stranded mRNA and TruSeq Stranded Total RNA Library Prep Kits. Version 2 contains improvements for 8 of the 192 index oligos in plate 20022371, with no changes to the remaining 184. Purchase library preps and probe panels separately.
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Illumina Ribo-Zero plus rRNA Depletion Kit (96 Samples)
20037135
Includes Ribo-Zero Plus depletion module for depletion of ribosomal RNA from human, mouse, rat, and bacteria and globin transcripts for 96 samples. Purchase purification beads, library prep reagents and index adapters separately. For complete kit, including library prep reagents, order the Illumina Stranded Total RNA with Ribo-Zero Plus kit.
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TruSeq Total RNA Training
15044069
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Limited Use Label License: This product and its use are the subject of one or more issued and/or pending U.S. and foreign patent applications owned by Max Planck Gesellschaft, exclusively licensed to New England Biolabs, Inc. and sublicensed to Illumina, Inc. The purchase of this product from Illumina, Inc., its affiliates, or its authorized resellers and distributors conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product by the buyer (whether the buyer is an academic or for profit entity). The purchase of this product does not convey a license under any claims in the foregoing patents or patent applications direct to producing the product. The buyer cannot sell or otherwise transfer this product or its components to a third party or otherwise use the product for the following COMMERICAL PURPOSES: (I) use of the product or its components in manufacturing; or (2) use of the product or its components for therapeutic or prophylactic purposes in human or animals.
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